Dystrophin Diagnosis: Comparison of Dystrophin Abnormalities by Immunofluorescence and Immunoblot Analyses

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 86; no. 18; pp. 7154 - 7158
• Main Authors: Arahata, Kiichi, Hoffman, Eric P., Kunkel, Louis M., Ishiura, Shoichi, Tsukahara, Toshifumi, Ishihara, Tadayuki, Sunohara, Nobuhiko, Nonaka, Ikuya, Ozawa, Eijiro, Sugita, Hideo
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 01.09.1989; National Acad Sciences
• Subjects: Amino Acid Sequence; Biological and medical sciences; Biomarkers - analysis; Biopsies; Carrier State; Cell membranes; Diseases of striated muscles. Neuromuscular diseases; Duchenne muscular dystrophy; Dystrophin; Fluorescence; Fluorescent Antibody Technique; Fluorescent antibody techniques; Humans; Immunoblotting; Medical sciences; Molecular Sequence Data; Molecular weight; Muscle Proteins - analysis; Muscle Proteins - genetics; Muscles - pathology; Muscular disorders; Muscular dystrophies; Muscular Dystrophies - diagnosis; Muscular Dystrophies - genetics; Muscular Dystrophies - pathology; Neurology; Neuromuscular diseases; Neuromuscular Diseases - diagnosis; Neuromuscular Diseases - genetics; Neuromuscular Diseases - pathology; Human; Becker muscular dystrophy; Duchenne muscular dystrophy; Immunoblotting assay; Diagnosis; Immunofluorescence; Localization
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.86.18.7154

Immunoblot characterization and immunofluorescence localization of dystrophin are presented for 76 human patients with various neuromuscular diseases. Normal dystrophin (shown by immunoblotting) was invariably visualized as a continuous, peripheral membrane immunostaining of myofibers. Biochemical abnormalities of dystrophin (either lower or higher molecular weight dystrophin) resulted in patchy, discontinuous immunostaining, suggesting that the abnormal dystrophin proteins are not capable of creating a complete membrane cytoskeleton network. There was a very strong correlation of clinical diagnoses with the type of dystrophin abnormality; all Duchenne muscular dystrophy patient muscle contained no detectable dystrophin, Becker muscular dystrophy patient muscle had clearly abnormal dystrophin, and unrelated diseases showed normal dystrophin. However, a single patient of five carrying the diagnosis of Fukuyama dystrophy showed no detectable dystrophin and thus appeared to be a Duchenne dystrophy patient by the biochemical assays. We know of no other case of a patient with a disease thought to be unrelated to Duchenne/Becker dystrophy yet demonstrating dystrophin deficiency. Based on the data presented, we conclude that immunofluorescence is the best technique for the detection of female carriers of Duchenne dystrophy, whereas immunoblotting appears superior for the prognostic diagnosis of Becker muscular dystrophy.